Background Hepatocellular carcinoma (HCC) is a leading cause of cancer-related mortality, with limited curative options for advanced disease. Natural Killer (NK) cells are critical innate immune effectors, but their anti-tumor function is severely compromised by the immunosuppressive tumor immune microenvironment (TIME), particularly through transforming growth factor-beta (TGF-beta). This study investigates the pivotal role of TGF-beta signaling in modulating NK cell phenotypes and functions within the HCC TIME.Methods to comprehensively assess TGF-beta pathway activation and its impact on NK cells, tumor-infiltrating lymphocytes (TILs) and liver-infiltrating lymphocytes (LILs) were isolated from HCC patients undergoing curative resection. Phenotypic and functional analyses were performed, along with functional restoration experiments targeting TGF-beta signaling.Results Tumor-infiltrating NK cells (TINKs) exhibited significant activation of both canonical (SMAD-dependent) and non-canonical (TAK1/p38 MAPK) TGF-beta signaling, with a predominance of the non-canonical pathway. This activation was associated with the emergence of an ILC1-like NK subset (CD103+/CD49a+), which was nearly absent in non-tumor liver tissue. These ILC1-like cells maintained strong cytokine production and expressed high levels of inhibitory receptors (PD-1, TIM-3, TIGIT), whereas conventional NK cells (cNKs; CD103-/CD49a-/CD9-) were functionally impaired. Notably, blocking TGF-beta receptor binding and SMAD3 activation restored cNK functionality.Discussion our findings suggest that while non-canonical TGF-beta signaling drives phenotypic reprogramming and contributes to NK cell dysfunction, canonical SMAD-dependent signaling remains a key therapeutic target for functional restoration. These results highlight the dual role of TGF-beta in immune modulation and suggest that targeted pathway inhibition could enhance innate anti-tumor responses, opening new avenues for combination therapies in HCC.
TGF-β-driven NK Cells plasticity in hepatocellular carcinoma / Reverberi, V.; Montali, A.; Vecchi, A.; Rossi, M.; Pelagatti, A.; Doselli, S.; Farina, B.; Olivani, A.; Economopoulos, G.; Dalla Valle, R.; Laccabue, D.; Ferraglia, F.; Penna, A.; Fisicaro, P.; Boni, C.; Missale, G.. - In: FRONTIERS IN IMMUNOLOGY. - ISSN 1664-3224. - 16:(2025). [10.3389/fimmu.2025.1651129]
TGF-β-driven NK Cells plasticity in hepatocellular carcinoma
Reverberi V.;Montali A.;Rossi M.;Pelagatti A.;Doselli S.;Farina B.;Dalla Valle R.;Laccabue D.;Boni C.
;Missale G.
2025-01-01
Abstract
Background Hepatocellular carcinoma (HCC) is a leading cause of cancer-related mortality, with limited curative options for advanced disease. Natural Killer (NK) cells are critical innate immune effectors, but their anti-tumor function is severely compromised by the immunosuppressive tumor immune microenvironment (TIME), particularly through transforming growth factor-beta (TGF-beta). This study investigates the pivotal role of TGF-beta signaling in modulating NK cell phenotypes and functions within the HCC TIME.Methods to comprehensively assess TGF-beta pathway activation and its impact on NK cells, tumor-infiltrating lymphocytes (TILs) and liver-infiltrating lymphocytes (LILs) were isolated from HCC patients undergoing curative resection. Phenotypic and functional analyses were performed, along with functional restoration experiments targeting TGF-beta signaling.Results Tumor-infiltrating NK cells (TINKs) exhibited significant activation of both canonical (SMAD-dependent) and non-canonical (TAK1/p38 MAPK) TGF-beta signaling, with a predominance of the non-canonical pathway. This activation was associated with the emergence of an ILC1-like NK subset (CD103+/CD49a+), which was nearly absent in non-tumor liver tissue. These ILC1-like cells maintained strong cytokine production and expressed high levels of inhibitory receptors (PD-1, TIM-3, TIGIT), whereas conventional NK cells (cNKs; CD103-/CD49a-/CD9-) were functionally impaired. Notably, blocking TGF-beta receptor binding and SMAD3 activation restored cNK functionality.Discussion our findings suggest that while non-canonical TGF-beta signaling drives phenotypic reprogramming and contributes to NK cell dysfunction, canonical SMAD-dependent signaling remains a key therapeutic target for functional restoration. These results highlight the dual role of TGF-beta in immune modulation and suggest that targeted pathway inhibition could enhance innate anti-tumor responses, opening new avenues for combination therapies in HCC.I documenti in IRIS sono protetti da copyright e tutti i diritti sono riservati, salvo diversa indicazione.
